Scientific Journal Impact Factor: 3.76 e- ISSN: -553 p- ISSN: -83X International Journal of Applied And Pure Science and Agriculture www.ijapsa.com ROLE OF MICROBIAL BEHAVIOR IN EXTRACT OF CLEOME GYNANDRA L. M.D. SARAVANAMOORTHY, R.SIVASANKARI, A. ZAHIRA3, S. KARTHIKA4 Assistant Professor of Botany, Arignar Anna Government Arts College, Musiri, Tiruchirappalli Dist, Tamil Nadu. India Assistant Professor in PG Department of Microbiology, SrimathiIndra Gandhi College, Tiruchirappalli, Tamil Nadu, India 3,4 Research Scholars in Zoology and Botany Department Arignar Anna Govt. Arts College, Musiri, Musi Tiruchirappalli Dist., Tamil Nadu. * Corresponding Author : M.D. SARAVANAMOORTHY Abstract : The study of solvent extracts on plant parts like root, stem and leaves of Cleome gynandra. gynandra L was studied against the reference bacterial strains of Escherichia coli, Enterobacterfaecalis, terfaecalis, and Proteus vulgaris.. The inhibition of bacterial strains was more pronounced with leaf solvent extract than stem and root. The percentage of inhibition was higher with E.coli and P.vulgaris of leaf solvent than compare to E. faecalis. Key words: Cleome gynandra L.; solvent extract; E.coli; Medicinal plant; Antibacterial activity. I. Introduction Antimicrobial properties of medicinal plants are being increasingly reported from different parts of the world?[]. Plants are indeed the first material used in alternative medicine type of remedy against many diseases []. Researchers are increasingly turning their attention to medicinal plant species looking to develop better antimicrobial drugs against infectious diseases. Several plants have therapeutic and pharmaceutical effects, for antimicrobial, antioxidant, anti anti-infectious ectious and antitumour activities 3,4]. Herbal medicine has been widely used as an integral part of primary health care in many countries3. Medicinal plants may constitute a reservoir of new antimicrobial substances to be discovered.a discovered. number of studies havee been reported, dealing with antimicrobial screening of extracts of medicinal plants [5],[3],[4],[5]&[6]. One of the medicinal plants known for having many medicinal uses in traditional system of medicine is Cleome gynandra gynandral. Due to its medicinal properties ies it cures a number of diseases such as head ache, piles, cough, muscular pain, eye complaints, cancer and skin diseases. However the plant C.gynandra L. has not been much scientifically investigated for its antibacterial activity [7]. Given the alarming incidence of antibiotic resistance in bacteria and fungi, there is a need therefore to identify new lead molecules, which could be exploited for developing ideal antimicrobial formulations [8]. In view of this the antibacterial activity of leaves, stem and root extracts of C.gynandra L. was studied In vitro against E.coli, Enterobacterfaecalis and Proteus vulgaris Many bacteria are naturally resistant to antibiotics due to the permeability barrier afforded by the outer membrane lipopolysaccharides ccharides (LPS). Also its tendency to colonize surface in a biofilm form makes the cells impervious to therapeutic concentration of antibiotics [9]. @IJAPSA-06, 06, All rights Reserved 67
Volume 0, Issue, [January - 06] e-issn: -553, p-issn: -83X Plant based medicine are more effective cheaper and alternative was probably responsible for the fast growing industry of herbal medicine, [0]. However due to over exploitation, some traditionally used plants are disappearing and the sustainable usage of natural resources is currently questioned by ecologists [& ]. To valid the traditional uses, the present study was under taken to determine the antibacterial activity of the same plant against some pathogenic bacteria. II. Materials and methods Plant material: The plants were collected from various location around Tiruchirappalli.. The plant materials (leaves stem and root) were dried under shade condition and ground well for powdered form. It was sealed with in polythene bags until the time of extraction. Preparation of the Extract: Five grams of fresh plant materials (leaves, stem and root) were extracted with 0 ml of distilled water, ethanol, methanol, petroleum ether and chloroform, They were kept for seven days at room temperature (3ºC) for complete extraction. After seven days the extracts were filtered through whatman No. filter paper. This extract was collected in bottles and kept in refrigerator for preservation. Bacterial Strains: The test organisms are mainly used for the antibacterial studies. They were identified by standard methods, and reference strains. They were preserved by frozen at 70o C. The following three bacterial strains were taken for antibacterial studies. Escherichia coli Enterobacterfaecalis and Proteus vulgaristhere were obtained from the microbial type culture collections. Bacterial strains were o maintained on nutrient agar slants at 4 C. Bacterial strains were subculture in liquid medium (Nutrient broth) at 370C for 8 hrs they were also used for subsequent tests. Methodology : Nutrient agar medium (ph. 7) is one of the most commonly used medium for several routine bacteriological purposes. After adding all the ingredients in to the distilled water it was boiled to dissolve the medium completely and was sterilized by autoclaving at 5 Psi pressure (oc) for 5 minutes. For the preparation of nutrient broth medium (ph. 7), after adding all the ingredients without agar in to the distilled water it was boiled to dissolve the medium completely and sterilized by autoclaving at 5 Lbs pressure (oc) for 5 minutes. Evaluationof the Antibacterial activity: Bacterial cultures (E.coli, E.faecalis andp.vulgaris) were maintained on nutrient agar at 4oC.They were obtained from the microbial type culture, Chandigarh (India). These bacterial cultures were diluted using nutrient broth and diluted bacterial cultures (0.ml) were spread over sterile Nutrient Agar plates, 0.ml of the plant extracts were applied per sterile filter paper disc (Whatman No..6 mm in diameter). The discs were allowed to dry before being placed to agar plates. Each plate contained three paper discs with plant extract. Each extract was tested in triplicate. The plates were incubated at 37oC for 4 hrs then the inhibition zones were recorded [] @IJAPSA-06, All rights Reserved 68
Volume 0, Issue, [January - 06] e-issn: -553, p-issn: -83X The resultant clear zones around the discs were measures (in cm) as a semi-quantitative indication of toxicity. The antibacterial activity of plant extracts were indicated by clear zones of growth inhibition. Random sampling method was used for the entire test in triplicates. Mean value and standard deviations was calculated using the formula given [3]. III. Results and Discussion The antibacterial sensitivity of the Cleomegynandra.L (leaf, stem and root) plant extracts and their potency was assessed using the disc diffusion methods by measuring the diameter of growth inhibition zones (Table to 3).The results revealed that the leaf extracted with ethanol had very large zones of inhibition ranging from.3 cm and it also showed high degree of inhibition against E.coli and P.vulgaris and also it exhibited significant inhibition against E.faecalis. The methanol, chloroform and leaf extract showed moderate inhibition against E.coli, E.faecalisandP.vulgaris. In the aqueous extract of leaf, contradictory results were observed. These extracts exhibited less inhibition against E.coli E.faecalis and P.vulgaris The stem extracted with ethanol, methanol and chloroform showed moderate effect on E.coli, E.faecalis, and P.vulgaris. The stem extract of the stem showed less inhibition against these bacterial strains. The ethanolic root extract of Cleome gynandra.l showed moderate activity against E.coli and also these extract showed or exhibited low activity on Proteus vulgaris andenterobacterfaecalis. Actively low inhibition was identified from methanol, chloroform, petroleum ether and aqueous root extracts against these three bacterial strains. From the results it appears that the higher activity resides in ethanolic leaf extract of the plant. Since other extracts including chloroform, methanol, petroleum ether and aqueous extracts did not effectively inhibit the growth of the bacteria. In our experiments, the organic extracts exhibited better antibacterial activity than aqueous extract. The antibacterial activity of plant extracts was not due to one main active chemical constituent but due to the combined action of other compounds involved in it [4]. On the general note, the root extracts of the plants showed no antibacterial activity while the leaf extracts exhibited more therapeutic effect on the tested isolates[5]. This is because the chemical constituents responsible for the antibacterial activity aremore soluble in ethanol extracts. The greater resistance of gram negative plant extracts has been documented previously [6][7]&[8] However, the plant Cleome gynandra.l has not been much scientifically investigated for its antibacterial activity [9]. Further studies are necessary to confirm the activity of unknown biochemical properties. Bibliography [] David and Clark, 998. Antimicrobial properties of medicinal plants. Journal ofethnopharmacology65: 75-8. [] Akroum S., Satta, D. and Lalauoui, K., Antibacterial, antioxidant, cytotoxic activities and phytochemical screening of some Algerian plants, Euro. J. Sci. Res 009, 3(), 89-95. [3] AmjadK., Dhia, S.K. and Abeer, K., Genetic relationship among salvia species and antimicrobial activity of their crude extracts against pathogenic bacteria. Asian Jour, Plant Sci, 005, 4(5), 544-549. @IJAPSA-06, All rights Reserved 69
Volume 0, Issue, [January - 06] e-issn: -553, p-issn: -83X [4] AmjadK., Dhia, S.K. and Abeer, K., Genetic relationship among salvia species and antimicrobial activity of their crude extracts against pathogenic bacteria. Asian Jour, Plant Sci, 005, 4(5), 544-549. [5] Malcom, S.A andsofowora, E.A. 969. Antimicrobial activity of selected Nigerian folk remedies and their constituent plants. Journal of Nationalproducts.3(4): 5-57. [6] Bhakuni, D.S., Bitther,M., Marticorena,C., Silva,M and Weldt,E. 974. Screening of Chilean plants for antimicrobial activity. Journal of Natural products, 37(4): 6-63. [7] Moskalenko, S.A. 986. Preliminary screening of Far.easternethnomedicinal plants for antibacterial activity. Journal of Ethnopharmacology. 5(3): 3-59. [8] Gundidza,M., and Gaza,N. 993. Antimicrobial activity of Dalbergiamelanoxylonextracts. Journal of Ethnopharmacology40: 7-30. [9] Perumalsamy,R., Ignacimuthu,S., 000. Antibacterial activity of some folklore medicinal plants used by tribal in Western Ghat of India. Journal ofethnopharmacology. 69: 63-7. [0] Srinivasan, D.,Sangeeth, N.,Suresh, T and Perumalsamy, P.L. 00. Antimicrobial activity of certain Indian Medicinal plants used in Folklore medicine. Journal of Ethnopharmacology.74 (8): 0-03. [] Kirtikar, K.R andbasu, D.D. 975. Indian medicinal plants, Jayyed press, New Delhi. [] Cowan, M.M., 999. Plant products as antimicrobial agents, Clinical Microbiology Review, 564-58. [3] Aburjai,T., Darwish,M., Al-Khalil,S., Mahafzah,A., and Al-Abbadi,A. 00. Screening of antibiotic resistant inhibitors from local plant materials against two different strains of Pseudomonas aeruginosa, Journal of Ethnopharmacology, 76: 39-44. [4] Rojas, A., Hernandez.,Pereda-Miranda, R and Mata, R. 99. Screening for antimicrobial activity of crude drug extracts and pure natural products from Mexican medicinal plants. Journal of Ethnopharmacology. 35: 69-7. [5] Akharaiyi, F.C. 0. Antibacterial, Phytochemical and Antioxidant activities of Daturametel. International Journal of PharmTech Research., Vol.3, No., pp 478-483. [6] Nigg,H.N and Seigler, D.99. Phytochemical Resources for Medicine and Agriculture. New York, Plenum press, P.76-363. [7] Cragg, G.M., Boyd, M.R., Cardellina, J.H., Grever, M.R., Schepartz, S.A., and Snader, K.M. 993. The role of plants in the drug discovery programme of the U.S Manipulation, In : International Crop Science Madison, USA: Crop Science Society of America. P 78-96. [8] Rosoanaivo and Ratsimamanga-Urverge., 993. Biological evaluation of plants with reference to the Malagasy flora. Monograph for the Ifs-NAPRECA workshop on Bio-assays. Antananaviva, Madagascar. pp7-79. [9] Gupta, S.P. 977. Statistical methods, S.Chand and Co., New Delhi. [0] Bai, D. 990. Traditional Chinese material: A respect and prospect, PlantaMedica,56: 50. [] Paz,E.A.,Cerdeiras,M.P.,Fernandex,J.,Moyna,P., Soubes,M., Vazquez,A., Vero,S and Zunino,L. 995. Screening of Uruguayan medicinal plants for antimicrobial activity. Journal ofethnopharmacology. 45: 67-70. [] Vlietinck, J., Van Hoof., Totte, J., Lasure, A., VandenBerghe, D., Wangabo, C., Mvukiyumwani, J. 995. Screening of hundred Rwandese medicinal plants for antimicrobial and antiviral properties. Journal of Ethnopharmacology. 46: 347. [3] Kudi, A.C., Umoh, J.U., Eduvie, L.O., Gefu, J. 999. Screening of some Nigerian medicinal plants for antibacterial activity. Journal of Ethnopharmacology. 67: 5-8. Table : Antibacterial sensitivity of Cleome gynandraon E.coli Test Organism E. Coli Extraction @IJAPSA-06, All rights Reserved Leaf.3±0.00 0.7 ± 0.0 0.6±0.08 0.4±0.00 0.5±0.00 0.4±0.04 0.4±0.00 0.3±0.08 0.±0.00 0.4±0.08 0.3±0.03 0.±0.08 0.±0.00 70
Volume 0, Issue, [January - 06] e-issn: ISSN: -553, p-issn: -83X Figure igure : Effect of Cleome gynandraon E. coli 3.5 3 Zones of Inhibition (cm).5.5 0.5 0 Plant parts Table : Antibacterial sensitivity of Cleome gynandraon Proetus vulgaris Inhibition zone (cm) Test organism Extraction Ethonal Methonal Proteus vulgaris Petroleum Ether Leaf. ±0.00 0.7 ± 0.08 0.5 ± 0.00 0.4 ± 0.08 0.3 ± 0.08 0.4 ± 0.00 0.4 ± 0.00 0.3 ± 0.04 0. ± 0.08 0. ± 0.00 0.3 ± 0.00 0.3 ± 0.00 0. ± 0.00 0. ± 0.08 0. ± 0.08 Figure : Effect of Cleome gynandraon Proteus vulgaris 3.5 Zone of inhibition (cm) 3.5.5 0.5 0 Plant parts @IJAPSA-06, 06, All rights Reserved 7
Volume 0, Issue, [January - 06] e-issn: ISSN: -553, p-issn: -83X Table 3: Antibacterial sensitivity of Cleome gynandraon Enterobacterfaecalis Test Organism Extraction Enterobacterfaecalis 0.9±0.00 0.5±0.04 0.4±0.08 0.±0.00 Inhibition zone (cm) 0.4±0.08 0.3±0.04 0.±0.00 0.±0.00 0.3±0.04 0.±0.00 0.±0.08 0.±0.08 Figure3:: Effect of Cleome gynandraon Enteribacterfaecalis Zone of inhibition (cm).5.5 0.5 0 Plant parts @IJAPSA-06, 06, All rights Reserved 7